A lipid droplet-targetable and biothiol-sensitive fluorescent probe for the diagnosis of cancer cells/tissues.

Lipid droplets (LDs) have recently been reported as an attractive target for cancer diagnosis and treatment, owing to their special structure or microenvironment changes in cancer development and resistance. However, the relationship between the biothiol level of LDs and cancer is still poorly understood, partially owing to the absence of effective molecular tools. Herein, we developed a LD-targetable and biothiol-sensitive fluorescent probe, BTDA-RSS, by introducing 2,4-dinitrobenzenesulfonyl (DNBS) as the biothiol reaction group into a benzothiazolyl derivative. BTDA-RSS displayed a marked and rapid fluorescence turn-on response toward biothiols, due to the biothiol-triggered cleavage of DNBS to yield the highly fluorescent benzothiazolyl iminocoumarin BTDA. In addition, the probe shows significant LD-targetable ability, and has been applied for imaging endogenous/exogenous biothiol changes in LDs. Importantly, BTDA-RSS has successfully been utilized to distinguish cancerous cells/tissues from normal cells/tissues with excellent contrast. Surprisingly, we demonstrated for the first time the visualization of LD biothiols in surgical specimens from cancer patients, thereby holding great potential for the application of BTDA-RSS in the clinical diagnosis of human cancers.

Real-Time Monitoring Mitochondrial Viscosity during Mitophagy Using a Mitochondria-Immobilized Near-Infrared Aggregation-Induced Emission Probe.

Mitophagy plays a crucial role in maintaining intracellular homeostasis through the removal of dysfunctional mitochondria and recycling their constituents in a lysosome-degradative pathway, which leads to microenvironmental changes within mitochondria, such as the pH, viscosity, and polarity. However, most of the mitochondrial fluorescence viscosity probes only rely on electrostatic attraction and readily leak out from the mitochondria during mitophagy with a decreased membrane potential, thus easily leading to an inaccurate detection of viscosity changes. In this work, we report a mitochondria-immobilized NIR-emissive aggregation-induced emission (AIE) probe CS-Py-BC, which allows for an off-on fluorescence response to viscosity, thus enabling the real-time monitoring viscosity variation during mitophagy. This system consists of a cyanostilbene skeleton as the AIE active core and viscosity-sensitive unit, a pyridinium cation for the mitochondria-targeting group, and a benzyl chloride subunit that induces mitochondrial immobilization. As the viscosity increased from 0.903 cP (0% glycerol) to 965 cP (99% glycerol), CS-Py-BC exhibited an about 92-fold increase in fluorescence intensity at 650 nm, which might be attributed to the restriction of rotation and inhibition of twisted intramolecular charge transfer in a high viscosity system. We also revealed that CS-Py-BC could be well immobilized onto mitochondria, regardless of the mitochondrial membrane potential fluctuation. Most importantly, using CS-Py-BC, we have successfully visualized the increased mitochondrial viscosity during starvation or rapamycin-induced mitophagy in real time. All these features render CS-Py-BC a promising candidate to investigate mitophagy-associated dynamic physiological and pathological processes.

Monitoring of the decreased mitochondrial viscosity during heat stroke with a mitochondrial AIE probe.

Heat stroke is a fatal condition which usually results in central nervous system dysfunction, organism damage and even death. The relationship between heat stroke and mitochondria is still relatively unknown due to a lack of suitable tools. Herein, an aggregation-induced emission (AIE) probe CSP, by introducing a pyridinium cation as the mitochondria-targeted group to an AIE active core cyanostilbene skeleton, is highly sensitive to viscosity changes due to the restriction of intramolecular motion (RIM) and inhibition of twisted intramolecular charge transfer (TICT) in high-viscosity systems. As expected, with the viscosity increasing from 0.903 cP (0% glycerol) to 965 cP (99% glycerol), CSP exhibited a significant enhancement (more than 117-fold) in fluorescence intensity at 625 nm, with an excellent linear relationship between log I 625 nm and log η (R2 = 0.9869, slope as high as 0.6727). More importantly, using CSP we have successfully monitored the decreased mitochondrial viscosity during heat stroke for the first time. All these features render the probe a promising candidate for further understanding the mechanism underlying mitochondria-associated heat stroke.

An esterase-sensitive AIEgen probe targeting mitochondria and lipid droplets for assessing cell viability.

In order to conduct in-depth research on the mechanisms of cancer diagnosis and treatment, it is very important to develop fluorescent probes to study the interactions between different organelles and understand the relationship between various organelles and cell viability. However, the lack of fluorescent probes to visualize two or more targets has resulted in limited studies of intracellular interactions between different organelles. To this end, in this work, we developed a near-infrared (NIR) AIE probe with dual-color emission, NAP-Py-E, for mitochondria and lipid droplets imaging. The probe NAP-Py-E consists of lipophilic fraction, pyridine cation structure and esterase hydrolysis site. Interestingly, NAP-Py-E first targets mitochondria and emits red fluorescence; after partially hydrolyed by esterase in living cells, the hydrolysate NAP-Py accumulates in lipid droplets and emits green fluorescence. The probe has been successfully used to assess cell viability due to its dual-color emission and dual-organelle targeted changes.

Stereocontrolled addition of Grignard reagents to oxa-bridged benzazepines: highly efficient synthesis of functionalized benzazepine scaffolds.

An efficient and highly diastereoselective synthesis of 2-substituted benzo[b]azepin-5-ol via stereocontrolled addition of Grignard reagents to oxa-bridged benzazepines has been developed. The reaction proceeds efficiently starting from versatile skeletons with mild reaction conditions as well as simple operation. Furthermore, 2-substituted benzazepinones could been obtained by simple Dess-Martin oxidation in excellent yields.